Project Summary/Abstract Influenza is a highly contagious viral infectious diseases and causes 250,000-500,000 deaths and 3-5 million severe illnesses each year worldwide. Cellular immunity is crucial to eliminate virus-infected cells and promote recovery. Cellular immunity against conserved antigens can also confer protection against drifted influenza viruses. Current influenza vaccines mainly induce neutralizing antibodies against highly variable surface antigens and poorly induces cellular immunity. Adjuvants are promising to enhance influenza vaccine-induced humoral and cellular immune responses. Yet, current adjuvants approved to boost influenza vaccination mainly enhance humoral immune responses with little effects on cellular immunity. This project develops a physical radiofrequency (RF)-based adjuvant (RFA) with potent humoral and cellular adjuvant effects to boost influenza vaccination with minimal local, systemic or long-term side effects. Our preliminary studies identified potent humoral and cellular adjuvants effects of non-invasive RF treatment on several vaccine antigens, such as model antigen ovalbumin, recombinant hemagglutinin, and pandemic 2009 influenza H1N1 vaccine. The humoral RFA effects are comparable and cellular RFA effects are superior to the most potent adjuvant used in influenza vaccines. We hypothesize the novel RFA will significantly boost influenza vaccine-induced humoral and cellular immune responses, resulting in increased protection against vaccine and non-vaccine viral strains. Three specific aims are proposed: To optimize RFA effects and delineate adjuvantation mechanisms (specific aim 1); To explore RFA to boost seasonal and pre-pandemic influenza vaccination (specific aim 2); To develop RFA-based universal T cell vaccine (specific aim 3). Safety and potency of RFA will be first explored in murine models and then validated in miniature pigs. The success of this project will allow us to submit Investigational Device Exemption applications to explore RFA to boost influenza vaccination in clinics. Our ultimate goal is to develop a handheld RF device for convenient and cost-effective adjuvantation of influenza vaccination.